Identification of Apoptotic Bodies in Equine Semen

Identification of Apoptotic Bodies in Equine Semen

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a...

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Published in:Reproduction in domestic animals Vol. 49; no. 2; pp. 254 - 262
Main Authors: Caselles, AB, Miro‐Moran, A, Morillo Rodriguez, A, Gallardo Bolaños, JM, Ortega‐Ferrusola, C, Salido, GM, Peña, FJ, Tapia, JA, Aparicio, IM
Format: Journal Article
Language:English
Published: Germany Blackwell Publishing Ltd 01-04-2014
Subjects:
Adult
Animal reproduction
Animals
Apoptosis
Apoptosis - physiology
caspases
correlation
Cryopreservation - methods
Cryopreservation - veterinary
fluorescent dyes
Horses
Horses - physiology
Humans
Male
semen
Semen - physiology
Semen Analysis - veterinary
Semen Preservation - methods
Semen Preservation - veterinary
Sperm
Sperm Motility
spermatogenesis
spermatozoa
Spermatozoa - physiology
stallions
testes
viability
Western blotting
Young Adult
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Summary:Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process.
Bibliography:http://dx.doi.org/10.1111/rda.12264
ark:/67375/WNG-94GC74KP-P
Ministerio Ciencia e Innovación - No. BES-2008-002106
ArticleID:RDA12264
Figure S1 Representative image comparing the size between apoptotic bodies found in plasma seminal and cytoplasmic droplet attached to the tail of the sperm..
Junta de Extremadura-FEDER - No. GR10010; No. PCE1002; No. DE12006
istex:40F71F0D37D2C9BDAF9703A0F40B6778810DE99C
Ministerio de Ciencia e Innovación-FEDER - No. BFU2011-30261; No. AGL 2010-20758
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0936-6768
1439-0531
DOI:10.1111/rda.12264